SummaryClear-cell renal cell carcinoma (ccRCC) exhibits a broad range of metastatic phenotypes that have not been systematically studied to date. Here, we analyzed 575 primary and 335 metastatic biopsies across 100 patients with metastatic ccRCC, including two cases sampledat post-mortem. Metastatic competence was afforded by chromosome complexity, and we identify 9p loss as a highly selected event driving metastasis and ccRCC-related mortality (p = 0.0014). Distinct patterns of metastatic dissemination were observed, including rapid progression to multiple tissue sites seeded by primary tumors of monoclonal structure. By contrast, we observed attenuated progression in cases characterized by high primary tumor heterogeneity, with metastatic competence acquired gradually and initial progression to solitary metastasis. Finally, we observed early divergence of primitive ancestral clones and protracted latency of up to two decades as a feature of pancreatic metastases.

OBJECTIVE To evaluate the presence of human papillomavirus (HPV) infection, the methylation status in the promoter region of thrombospondin‐1 (TSP‐1), RAS association domain family 1A (RASSF1‐A) and p16 genes, and the expression of TSP‐1, CD31, p16 and p53 proteins in patients diagnosed with penile cancer, and the possible associations between these variables and clinical and pathological features. PATIENTS AND METHODS HPV types, gene promoter hypermethylation and protein expression were analysed by reverse line blot, methylation‐specific polymerase chain reaction, and immunohistochemistry, respectively, in 24 penile squamous cell carcinomas. RESULTS HPV infection was detected in 11 of 24 cases (46%), and TSP‐1, RASSF1‐A and p16 genes were hypermethylated in 46%, 42% and 38% of the tumours, respectively. TSP‐1 hypermethylation was associated with unfavourable histological grade (grade 3; P = 0.033), vascular invasion (P = 0.023), weak expression of TSP‐1 protein (P = 0.041), and shorter overall survival (P = 0.04). TSP‐1 expression was not associated with microvessel density. However, RASSF1‐A hypermethylation was more frequent in T1 tumours (P = 0.01), and p16 hypermethylation was not associated with any of the tested variables except for absence of p16 expression (P = 0.022). CONCLUSION In summary, the epigenetic inactivation of TSP‐1 and RASSF1‐A genes is associated with pathological variables and seems to be of prognostic significance in penile cancer.

Squamous cell carcinoma (SCC) of the vulva is a heterogeneous disease, associated or not with vulvar lichen sclerosus (LS). The precursor role of LS in vulvar cancer is unclear. We studied the epigenetic alterations of RASSF1A, RASSF2A, p16, TSP-1 and MGMT genes in vulvar SCCs, LS associated with SCC, isolated LS and normal vulvar skin. Gene hypermethylation and human papillomavirus presence were evaluated by methylation-specific PCR and PCR/reverse line blot, respectively. High-risk human papillomavirus types were present in 16.7% of the patients with vulvar SCC. There were increasing percentages of hypermethylation of genes from isolated LS to LS associated with vulvar SCC and vulvar SCC. The genes were hypermethylated more frequently in vulvar SCC associated with LS than in those not associated with LS, MGMT and RASSF2A being unmethylated in LS not associated with vulvar SCC. TSP-1 hypermethylation was related to recurrence in patients with vulvar cancer. Conclusions are as follows: (i) the epigenetic inactivation of genes is a common event in vulvar SCC and is also present in adjacent lesions, implying a possible precursor role for these alterations; (ii) MGMT and RASSF2A hypermethylation are present exclusively in vulvar SCC and LS associated with SCC, and absent from isolated LS; and (iii) TSP-1 hypermethylation is a bad prognosis factor in vulvar SCC.Vulvar cancer (VC) accounts for 3-5% of all female genital cancers and is more frequent in women older than 75 years. Squamous cell carcinoma (SCC) is the most common type of VC and has been divided into two groups on the basis of the presence or absence of the human papillomavirus (HPV), which leads to different models of progression.2 It is well known that there are, on one hand, HPV-associated VCs of the warty or basaloid type and, on the other, keratinizing SCC, most of which are not HPV related. First, SCC associated with HPV, mostly HPV16, is the least frequent, accounting for one third of vulvar SCC. It is diagnosed at young ages and is associated with other types of cancer, such as vaginal and anal carcinoma. It shares risk factors with cervical cancer (multiple sexual partners, previous history of smoking and HPV presence and typical pathological features with warty or basaloid subtypes). The precursor lesion is of the classic vulvar intraepithelial neoplasia (VIN) type, and 4% of VIN have a risk of malignancy and frequently relapse spontaneously. 4 On the other hand, SCC that is not associated with HPV is more frequent (66% of cases), typical of older ages and more frequently associated with lichen sclerosus (LS) or squamous cell hyperplasia. 4,5 The exact role of LS in pathogenesis of VC is not yet known, but the risk of developing vulvar SCC approaches 5% in women with these lesions.5 SCC is of the keratinizing type and is preceded by differentiated VIN, which is characterized by atypical basal keratinocytes with normal maturation. These precursor lesions are rarely diagnosed, their grade of malignancy is high and they exhibit immunohistochemical ...

Purpose:To analyze immunohistochemically morules in endometrioid lesions to show that CD10 is a sensitive marker for morular metaplasia. Experimental Design: Immunohistochemical analysis of 53 instances of morular metaplasia comprising 1 cyclic endometrium and 52 endometrioid lesions associated with focal glandular complexity corresponding to 9 polyps, 4 atypical polypoid adenomyomas, 24 complex endometrial hyperplasias (18 with and 6 without atypia), 12 grade 1 endometrioid adenocarcinomas in early clinical stages of both uterus and ovary, and three ovarian adenofibromas. Immunohistochemistry in paraffin sections was done for CD10, h-catenin, estrogen and progesterone receptors, and cytokeratins 5-6, 7, 8, 13, 18, 19, 20, and 34h-E12. Results: Morules were negative for estrogen and progesterone receptors and had h-cateninp ositive nuclei. Cytokeratins 8, 18, 19 were positive; cytokeratins 7 and 20 were negative; and cytokeratins 5-6, 13, and 34h-E12 were weakly positive. All cases revealed strongly positive membranous CD10 staining in morules, which was absent in glands. CD10 positivity allowed easy identification of morules at low power in various types of surgical specimens and in curettings. CD10 also highlighted early morular metaplasia in glandular epithelium. In cases associated with squamous, keratinizing metaplasia, CD10 discriminated between both types of metaplasia. Conclusions: CD10 staining represents a useful marker of morules in endometrioid neoplasms of the female genital tract, permitting identification of lesions usually associated with an attenuated malignancy. Considering the immunohistochemical and genetic similarities of morules in tumors of different organs, it is likely that this marker may be also useful to diagnose morular metaplasia in similar neoplasms of extragenital locations.Morules (1) are nodular structures found in endometrial-type glands formed by a peculiar metaplastic non-keratinizing squamoid epithelium (2). Morphologically similar structures have also been described in neoplastic lesions in other anatomic sites, such as thyroid (3), lung (4, 5), stomach (6), pancreas (7), and colon (8).In both eutopic and ectopic endometrioid tissues, morules are almost invariably associated with the glandular architectural complexity of premalignant and low-grade glandular malignant lesions but are absent in high-grade ones. Thus, it can be said that the presence of morules in the endometrium relates well with an attenuated malignancy.This study deals with the immunohistochemical findings in a series of 53 cases of morular metaplasia from both uterus and ovary, showing for the first time that CD10 staining is a marker of morules, allowing its easy identification in various endometrioid lesions. Materials and MethodsA total of 53 cases of endometrioid morular metaplasia were collected from the files of the

Mismatch repair deficiency (MMRD) is involved in the initiation of both hereditary and sporadic tumors. MMRD has been extensively studied in colorectal cancer and endometrial cancer, but not so in other tumors, such as ovarian carcinoma. We have determined the expression of mismatch repair proteins in a large cohort of 502 early-stage epithelial ovarian carcinoma entailing all the 5 main subtypes: high-grade serous carcinoma, endometrioid ovarian carcinoma (EOC), clear cell carcinoma (CCC), mucinous carcinoma, and low-grade serous carcinoma. We studied the association of MMRD with clinicopathologic and immunohistochemical features, including tumor-infiltrating lymphocytes in EOC, the histologic type in which MMRD is most frequent. In addition, MLH1 promoter methylation status and massive parallel sequencing were used to evaluate the proportion of sporadic and Lynch syndrome–associated tumors, and the most frequently mutated genes in MMRD EOCs. MMRD occurred only in endometriosis-associated histologic types, and it was much more frequent in EOC (18%) than in CCC (2%). The most frequent immunohistochemical pattern was loss of MLH1/PMS2, and in this group, 80% of the cases were sporadic and secondary to MLH1 promoter hypermethylation. The presence of somatic mutations in mismatch repair genes was the other mechanism of MMRD in sporadic tumors. In this series, the minimum estimated frequency of Lynch syndrome was 35% and it was due to germline mutations in MLH1, MSH2, and MSH6. ARID1A, PTEN, KTM2B, and PIK3CA were the most common mutated genes in this series. Interestingly, possible actionable mutations in ERRB2 were found in 5 tumors, but no TP53 mutations were detected. MMRD was associated with younger age and increased tumor-infiltrating lymphocytes. Universal screening in EOC and mixed EOC/CCC is recommended for the high frequency of MMRD detected; however, for CCC, additional clinical and pathologic criteria should be evaluated to help select cases for analysis.